目的：研究新型染色体区域稳定蛋白1（chromosome region maintenance 1, CRM1)抑制剂S109对非小细胞肺癌细胞增殖的影响及其机制。方法： S109作用于非小细胞肺癌细胞株H1975和H1650，以CCK-8法检测其对细胞生长的影响，以EdU法检测细胞中对细胞增殖的影响，以集落形成实验检测对细胞克隆形成能力的影响，以流式细胞术检测其对细胞周期的影响，以免疫荧光法检测对核输出标志蛋白RanBP1细胞内定位的影响，以Western blotting检测对细胞中CRM1和Cyclin D1蛋白表达的影响。结果： S109（0.04、0.2、1、5、25和50 μmol/L）可以剂量依赖方式显著抑制非小细胞肺癌H1975和H1650细胞生长（IC50值分别为1.4和1.2 μmol/L）；2和4 μmol/L的S109均可以显著抑制非小细胞肺癌增殖\[（51.3±28）%、（23.2±21）% vs（100±1.2）%, P<0.01\]和克隆形成\[（43.6±3.2）%、（26.8±2.8）% vs（100±1.4）%, P<001\]，并使细胞阻滞在G1期（G1期细胞数量由50.5%增加至74.9%）。S109可以特异性抑制核质转运标志蛋白RanBP1的核输出，并诱导CRM1蛋白降解。结论： S109通过特异性抑制CRM1功能阻断核输出，从而抑制非小细胞肺癌细胞增殖并阻滞细胞周期。

Objective:To investigate if and how chromosome region maintenance 1 (CRM1) inhibitor S109 affects the proliferation of human lung cancer cells in vitro. Methods: Non-small cell lung cancer (NSLC) H197 and H1650 cells were treated with S109 at 0.04, 0.2, 1, 5, 25, 50 μmol/L. After treatment for 72 h, cell viability, proliferation and colony formation were examined by CCK-8, EdU and clonogenic assays, respectively. Cell cycle arrest and subcellular localization of RanBP1 were analyzed by flow cytometry and fluorescence microscopy respectively. CRM1 and Cyclin D1 protein contents were assessed by Western blotting. Results: S109 inhibited the growth of H1975 and H1650 cells in a dose-dependent manner and the IC50 in these two cell lines was 1.4 and 1.2 μmol/L, respectively. Compared with control, S109 reduced H1975 cell proliferation by （51.3±2.8）% at 2 μmol/L and by (23.2±2.1)% at 4 μmol/L (P<0.05) and reduced colony formation by (43.6±3.2)% at 2 μmol/L and by (26.8±2.8)% at 4 μmol/L (P<0.05). S109 treatment increased the G1 fraction from 50.5% to 74.9% in H1975 cells (P<0.05), specifically inhibited nuclear export of RanBP1 and induced degradation of CRM1. Conclusion: S109 is able to suppress the proliferation and induces cell cycle arrest of NSLC cells, at least partially, via inhibiting nuclear export of RanBP1.

国家自然科学基金资助项目（No. 81400167, No.81402074）